Objective:To investigate the anti-melanogenic potential of ligustroside isolated from Ligustrum japonicum.Methods:The cytotoxicity of ligustroside was tested via MTT assay.Furthermore,the effects of ligustroside on th...Objective:To investigate the anti-melanogenic potential of ligustroside isolated from Ligustrum japonicum.Methods:The cytotoxicity of ligustroside was tested via MTT assay.Furthermore,the effects of ligustroside on the expression of critical melanogenic markers such as tyrosinase,tyrosinase related proteins(TRPs),and microphthalmia-associated transcription factor(MITF)were analyzed at both mRNA and protein levels via RT-qPCR and Western blot,respectively,inα-melanocyte stimulating hormoneinduced B16F10 cells.In addition,phosphorylation of p38,ERK and JNK proteins was investigated.Immunofluorescence analysis of MITF was also conducted.Results:Ligustroside significantly reduced intracellular tyrosinase activity and melanin content by 37.11%and 29.12%,respectively,compared to untreated cells.Moreover,it downregulated the expression of MITF,tyrosinase,TRP-1,and TRP-2 at the mRNA and protein levels by regulating both the mitogen-activated protein kinase(MAPK)and protein kinase A(PKA)/cAMP response element-binding protein(CREB)signaling pathways.Ligustroside also suppressed the nuclear protein expression of MITF,β-catenin,and p-CREB,and decreased immunofluorescence intensity of nuclear MITF.Conclusions:Ligustroside derived from Ligustrum japonicum shows a significant anti-melanogenesis effect via suppression of the MAPK and PKA/CREB signaling pathways.展开更多
Hyperlipidemia is a risk factor for clinically significant thrombotic events in cardiovascular diseases.Platelet reactivity in hyperlipidemic conditions is enhanced when platelet scavenger receptor CD36 recognizes oxi...Hyperlipidemia is a risk factor for clinically significant thrombotic events in cardiovascular diseases.Platelet reactivity in hyperlipidemic conditions is enhanced when platelet scavenger receptor CD36 recognizes oxidized lipids in oxidized low-density lipoprotein(ox-LDL)particles,a process that induces atherothrombosis.Sulforaphane(SFN)is a dietary isothiocyanate enriched in cruciferous vegetables and exerts multiple biological activities.The current study sought to investigate the efficacy of SFN on platelet hyperreactivity under hyperlipidemic conditions in vitro and in vivo.Using a series of platelet functional assays in human platelets in vitro,we demonstrated that SFN attenuated ox-LDL-increased platelet aggregation and activation(surface CD62P expression).Mechanistically,studies using pharmacological inhibitors clarified that these inhibitory effects of SFN were mainly modulated by down-regulating CD36-mediated activation of Src kinases,leading to enhanced activation of cyclic adenosine monophosphate/protein kinase A(cAMP/PKA)signaling,and resultant inhibition of NADPH oxidase 2(NOX2)-dependent generation of reactive oxygen species(ROS).Moreover,12-week supplementation of SFN-enriched broccoli sprout extract(BSE,0.06%diet)in hyperlipidemic C57BL/6J mice also decreased platelet hyperreactivity.Studies using pharmacological inhibitors of CD36,protein kinase A(PKA)and NOX2 showed that the efficacy of BSE supplementation was mainly through modulating CD36-mediated the cAMP/PKA/NOX2 signaling.Thus,through modulating the cAMP/PKA/NOX2 pathway and attenuating CD36-mediated platelet hyperreactivity,SFN may play important protective roles in atherothrombosis under hyperlipidemic conditions.展开更多
基金supported by the BB21plus funded by Busan Metropolitan City and Busan Techno Park,and the National Research Foundation of Korea(NRF)grant funded by the Korea government(MSIT)(No.NRF-2023R1A2C1006268 and RS-2023-00212560).
文摘Objective:To investigate the anti-melanogenic potential of ligustroside isolated from Ligustrum japonicum.Methods:The cytotoxicity of ligustroside was tested via MTT assay.Furthermore,the effects of ligustroside on the expression of critical melanogenic markers such as tyrosinase,tyrosinase related proteins(TRPs),and microphthalmia-associated transcription factor(MITF)were analyzed at both mRNA and protein levels via RT-qPCR and Western blot,respectively,inα-melanocyte stimulating hormoneinduced B16F10 cells.In addition,phosphorylation of p38,ERK and JNK proteins was investigated.Immunofluorescence analysis of MITF was also conducted.Results:Ligustroside significantly reduced intracellular tyrosinase activity and melanin content by 37.11%and 29.12%,respectively,compared to untreated cells.Moreover,it downregulated the expression of MITF,tyrosinase,TRP-1,and TRP-2 at the mRNA and protein levels by regulating both the mitogen-activated protein kinase(MAPK)and protein kinase A(PKA)/cAMP response element-binding protein(CREB)signaling pathways.Ligustroside also suppressed the nuclear protein expression of MITF,β-catenin,and p-CREB,and decreased immunofluorescence intensity of nuclear MITF.Conclusions:Ligustroside derived from Ligustrum japonicum shows a significant anti-melanogenesis effect via suppression of the MAPK and PKA/CREB signaling pathways.
基金supported by the National Natural Science Foundation of China(82003451 and 82003455)Yunnan Fundamental Research Projects(202101AT070033)the Start-Up Fund for Introduction of High-level Talents to Dali University(YBS2021015).
文摘Hyperlipidemia is a risk factor for clinically significant thrombotic events in cardiovascular diseases.Platelet reactivity in hyperlipidemic conditions is enhanced when platelet scavenger receptor CD36 recognizes oxidized lipids in oxidized low-density lipoprotein(ox-LDL)particles,a process that induces atherothrombosis.Sulforaphane(SFN)is a dietary isothiocyanate enriched in cruciferous vegetables and exerts multiple biological activities.The current study sought to investigate the efficacy of SFN on platelet hyperreactivity under hyperlipidemic conditions in vitro and in vivo.Using a series of platelet functional assays in human platelets in vitro,we demonstrated that SFN attenuated ox-LDL-increased platelet aggregation and activation(surface CD62P expression).Mechanistically,studies using pharmacological inhibitors clarified that these inhibitory effects of SFN were mainly modulated by down-regulating CD36-mediated activation of Src kinases,leading to enhanced activation of cyclic adenosine monophosphate/protein kinase A(cAMP/PKA)signaling,and resultant inhibition of NADPH oxidase 2(NOX2)-dependent generation of reactive oxygen species(ROS).Moreover,12-week supplementation of SFN-enriched broccoli sprout extract(BSE,0.06%diet)in hyperlipidemic C57BL/6J mice also decreased platelet hyperreactivity.Studies using pharmacological inhibitors of CD36,protein kinase A(PKA)and NOX2 showed that the efficacy of BSE supplementation was mainly through modulating CD36-mediated the cAMP/PKA/NOX2 signaling.Thus,through modulating the cAMP/PKA/NOX2 pathway and attenuating CD36-mediated platelet hyperreactivity,SFN may play important protective roles in atherothrombosis under hyperlipidemic conditions.
